Molecular characterization of two candidate genes associated with coat color in Tibetan sheep (Ovis arise)
HAN Ji-long, YANG Min, GUO Ting-ting, YUE Yao-jing, LIU Jian-bin, NIU Chun-e, WANG Chao-feng, YANG Bo-hui
1、Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050,P.R.China
2、Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
3、College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.China
摘要 Coat color is a key economic trait in sheep. Some candidate genes associated with animal’s coat color were found. Particularly, v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) and microphthalmia-associated transcription factor (MITF) play a key role in the modulation of hair pigmentation in mammals. This study investigated those two candidate genes’ mutations and expressions associated with wool color in Tibetan sheep. First, the gene polymorphisms of those two genes were analyzed, and then, relative mRNA expression levels of those two genes in skin tissue with different coat colors were compared. Thirdly, KIT and MITF protein expression levels were detected through Western blot and immunehistochemical. Allele C was predominant allele in the white coat color Tibetan sheep population of the MITF coding region g. 1548 C/T loci. The relative MITF mRNA expression in black coat skin tissue was significantly higher than white (P<0.01). However, no significant differences were detected in the KIT gene’s mRNA expression of these two different coat color skin tissues (P>0.05), while the level of KIT protein expression in skin tissues of white and black coats was also roughly equivalent. Our study observed that, the level of MITF protein expression in black coat skin tissue was significantly higher than that in white coat skin tissue, and positive staining for MITF protein expression was detected mainly in the epidermis and the dermal papilla, bulb, and outer root sheath of hair follicles. We conclude that the black coat of Tibetan sheep is related to high MITF expression in the hair follicles, and MITF may be important for coat color formation of Tibetan sheep.
Abstract Coat color is a key economic trait in sheep. Some candidate genes associated with animal’s coat color were found. Particularly, v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) and microphthalmia-associated transcription factor (MITF) play a key role in the modulation of hair pigmentation in mammals. This study investigated those two candidate genes’ mutations and expressions associated with wool color in Tibetan sheep. First, the gene polymorphisms of those two genes were analyzed, and then, relative mRNA expression levels of those two genes in skin tissue with different coat colors were compared. Thirdly, KIT and MITF protein expression levels were detected through Western blot and immunehistochemical. Allele C was predominant allele in the white coat color Tibetan sheep population of the MITF coding region g. 1548 C/T loci. The relative MITF mRNA expression in black coat skin tissue was significantly higher than white (P<0.01). However, no significant differences were detected in the KIT gene’s mRNA expression of these two different coat color skin tissues (P>0.05), while the level of KIT protein expression in skin tissues of white and black coats was also roughly equivalent. Our study observed that, the level of MITF protein expression in black coat skin tissue was significantly higher than that in white coat skin tissue, and positive staining for MITF protein expression was detected mainly in the epidermis and the dermal papilla, bulb, and outer root sheath of hair follicles. We conclude that the black coat of Tibetan sheep is related to high MITF expression in the hair follicles, and MITF may be important for coat color formation of Tibetan sheep.
HAN Ji-long, YANG Min, GUO Ting-ting, YUE Yao-jing, LIU Jian-bin, NIU Chun-e, WANG Chao-feng, YANG Bo-hui.
2015.
Molecular characterization of two candidate genes associated with coat color in Tibetan sheep (Ovis arise). Journal of Integrative Agriculture, 14(7): 1390-1397.
Bai D, Yang L, Unerhu, Zhao Y, Zhao Q, Gaowa H, Manglai D.2011. Effects of KIT gene on coat depigmentation in whitehorses. Hereditas (Beijing), 33, 1171-1178
Bertolotto C, Abbe P, Hemesath T J, Bille K, Fisher D E,Ortonne J P, Ballotti R 1998. Microphthalmia gene productas a signal transducer in cAMP-induced differentiation ofmelanocytes. Journal of Cell Biology, 142, 827-835
Dong C S, Wang H D, Xue L L, Dong Y J, Yang L, Fan R W,Yu X J, Tian X, Ma S H, Smith G W. 2012. Coat colordetermination by miR-137 mediated down-regulation ofmicrophthalmia-associated transcription factor in a mousemodel. RNA, 18, 1679-1686
Fleet M R. 2002. Pigmentation prevention in sheep: Complexor simple? Wool Technology and Sheep Breeding, 50,410-416
Fontanesi L, Scotti E, Russo V. 2012. Haplotype variability inthe bovine MITF gene and association with piebaldism inHolstein and Simmental cattle breeds. Animal Genetics,43, 250-256
Gundry C N, Dobrowolski S F, Martin Y R, Robbins T C, NayL M, Boyd N, Coyne T, Wall M D, Wittwer C T, Teng D.2008. Base-pair neutral homozygotes can be discriminatedby calibrated high-resolution melting of small amplicons.Nucleic Acids Research, 36, 3401-3408
Haase B, Brooks S A, Tozaki T, Burger D, Poncet P A, RiederS, Hasegawa T, Penedo C, Leeb T. 2009. Seven novelKIT mutations in horses with white coat colour phenotypes.Animal Genetics, 40, 623-629
Hauswirth R, Haase B, Blatter M, Brooks S A, Burger D,Drogemuller C, Gerber V, Henke D, Janda J, Jude R. 2012.Mutations in MITF and PAX3 cause “splashed white” andother white spotting phenotypes in horses. Plos Genetics,8, 404-412
Hayes B J, Pryce J, Chamberlain A J, Bowman P J, Goddard ME. 2010. Genetic architecture of complex traits and accuracyof genomic prediction: coat colour, milk-fat percentage,and type in Holstein cattle as contrasting model traits. PlosGenetics, 6, e1001139.
Hemesath T J, Steingrimsson E, McGill G, Hansen M J, VaughtJ, Hodgkinson C A, Arnheiter H, Copeland N G, JenkinsN A, Fisher D E. 1994. Microphthalmia, a critical factor inmelanocyte development, defines a discrete transcriptionfactor family. Genes and Development, 8, 2770-2780
Horikawa T, Norris D A, Johnson T W, Zekman T, DunscombN, Bennion S D, Jackson R L, Morelli J G. 1996. DOPAnegativemelanocytes in the outer root sheath of humanhair follicles express premelanosomal antigens but nota melanosomal antigen or the melanosome-associatedglycoproteins tyrosinase, TRP-1, and TRP-2. Journal ofInvestigative Dermatology, 106, 28-35
Kadekaro A L, Kavanagh R J, Wakamatsu K, Ito S, PipitoneM A, Abdel-Malek Z A. 2003. Cutaneous photobiology.The melanocyte vs. the sun: Who will win the final round?Pigment Cell Research, 16, 434-447
Kijas J W, Lenstra J A, Hayes B, Boitard S, Neto L, SanCristobal M, Servin B, McCulloch R, Whan V, Gietzen K.2012. Genome-wide analysis of the world’s sheep breedsreveals high levels of historic mixture and strong recentselection. Plos Biology, 10, e1001258.
Lalwani A K, Attaie A, Randolph T, Deshmukh D, Wang C,Mhatre A, Wilcox E. 1998. Point mutation in the MITF gene causing Waardenburg syndrome type II in a threegenerationIndian family. American Journal of MedicalGenetics, 80, 406-409
Lee H D, Lee W H, Roh E, Seo C S, Son J K, Lee S H, Hwang BY, Jung S H, Han S B, Kim Y. 2011. Manassantin A inhibitscAMP-induced melanin production by down-regulating thegene expressions of MITF and tyrosinase in melanocytes.Experimental Dermatology, 20, 761-763
Li J Y, Song J S, Bell R, Tran T, Haq R, Liu H F, Love K T, LangerR, Anderson D G, Larue L, Fisher D E. 2012. YY1 regulatesmelanocyte development and function by cooperating withMITF. PLoS Genetics, 8, e1002688.
Limat A, Salomon D, Carraux P, Saurat J H, Hunziker T.1999. Human melanocytes grown in epidermal equivalentstransfer their melanin to follicular outer root sheathkeratinocytes. Archives of Dermatological Research, 291,325-332
Lin J Y, Fisher D E. 2007. Melanocyte biology and skinpigmentation. Nature, 445, 843-850
Lister J A, Robertson C P, Lepage T, Johnson S L, Raible D W.1999. Nacre encodes a zebrafish microphthalmia-relatedprotein that regulates neural crest-derived pigment cell fate.Development, 126, 3757-3767
Liu L, Harris B, Keehan M, Zhang Y. 2009. Genome scan forthe degree of white spotting in dairy cattle. Animal Genetics,40, 975-977
MacKenzie M, Jordan S A, Budd P S, Jackson I J. 1997.Activation of the receptor tyrosine kinase Kit is requiredfor the proliferation of melanoblasts in the mouse embryo.Developmental Biology, 192, 99-107
Nakayama A, Nguyen M, Chen C C, Opdecamp K, HodgkinsonC A, Amheiter H. 1998. Mutations in microphthalmia, themouse homolog of the human deafness gene MITF, affectneuroepithelial and neural crest-derived melanocytesdifferently. Mechanisms of Development, 70, 155-166
Opdecamp K, Vanvooren P, Riviere M, Arnheiter H, MottaR, Szpirer J, Szpirer C. 1998. The rat microphthalmiaassociatedtranscription factor gene (Mitf) maps at 4q34-q41and is mutated in the mib rats. Mammalian Genome, 9,617-621
Otreba M, Rok J, Buszman E, Wrzesniok D. 2012. Regulationof melanogenesis: the role of cAMP and MITF. PostepyHigieny I Medycyny Doswladczalnej, 66, 33-40
(in Polish)Penagaricano F, Zorrilla P, Naya H, Robello C, Urioste J I. 2012.Gene expression analysis identifies new candidate genesassociated with the development of black skin spots inCorriedale sheep. Journal of Applied Genetics, 53, 99-106
Peters E, Tobin D J, Botchkareva N, Maurer M, Paus R. 2002.Migration of melanoblasts into the developing murine hairfollicle is accompanied by transient c-Kit expression. Journalof Histochemistry & Cytochemistry, 50, 751-766
Philipp U, Lupp B, Momke S, Stein V, Tipold A, Eule J C,Rehage J, Distl O. 2011. A MITF mutation associated with adominant white phenotype and bilateral deafness in GermanFleckvieh cattle. PLoS One, 6, e28857.
Royo L J, Alvarez I, Arranz J J, Fernandez I, Rodriguez A,Perez-Pardal L, Goyache F. 2008. Differences in theexpression of the ASIP gene are involved in the recessiveblack coat colour pattern in sheep: evidence from the rareXalda sheep breed. Animal Genetics, 39, 290-293
Shoag J, Haq R, Zhang M, Liu L, Rowe G C, Jiang A, KoulisisN, Farrel C, Amos C I, Wei Q. 2013. PGC-1 coactivatorsregulate MITF and the tanning response. Molecular Cell,49, 145-157
Steingrimsson E, Copeland N G, Jenkins N A. 2004.Melanocytes and the microphthalmia transcription factornetwork. Annual Review of Genetics, 38, 365-411
Takeda K, Yasumoto K, Kawaguchi N, Udono T, WatanabeK, Saito H, Takahashi K, Noda M, Shibahara S. 2002.Mitf-D, a newly identified isoform, expressed in the retinalpigment epithelium and monocyte-lineage cells affectedby Mitf mutations. Biochimica et Biophysica Acta-GeneStructure and Expression, 1574, 15-23
Taylor B A, Navin A, Phillips S J. 1994. PCR-Amplification ofsimple sequence repeat variants from pooled DNA samplesfor rapidly mapping new mutations of the mouse. Genomics,21, 626-632
Vachtenheim J, Borovansky J. 2010. “Transcription physiology”of pigment formation in melanocytes: central role of MITF.Experimental Dermatology, 19, 617-627
Yasumoto K, Amae S, Udono T, Fuse N, Takeda K, ShibaharaS. 1998. A big gene linked to small eyes encodes multipleMitf isoforms: Many promoters make light work. PigmentCell Research, 11, 329-336
Zhang R, Fan R, Cheng Z, Tian X, Liu J, Gao L, Ma Z, Dong C.2011. Regulation of TYR and MITF mRNA expression byCDK5 in alpaca melanocytes. Chinese Journal of Animaland Veterinary Sciences, 42, 1712-1717 (in Chinese)
Zhu Z, He J, Jia X, Jiang J, Bai R, Yu X, Lv L, Fan R, He X,Geng J. 2010. MicroRNA-25 functions in regulation ofpigmentation by targeting the transcription factor MITF inalpaca (Lama pacos) skin melanocytes. Domestic AnimalEndocrinology, 38, 200-209
Atiqur RAHMAN, Md. Hasan Sofiur RAHMAN, Md. Shakil UDDIN, Naima SULTANA, Shirin AKHTER, Ujjal Kumar NATH, Shamsun Nahar BEGUM, Md. Mazadul ISLAM, Afroz NAZNIN, Md. Nurul AMIN, Sharif AHMED, Akbar HOSAIN. Advances in DNA methylation and its role in cytoplasmic male sterility in higher plants[J]. >Journal of Integrative Agriculture, 2024, 23(1): 1-19.
[4]
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